Re: Stray light in TIRF microscopy

Posted by John Oreopoulos on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Stray-light-in-TIRF-microscopy-tp7585414p7585427.html

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Dear Alexander,

I observed the stray light phenomenon for through-the objective TIRF in this paper:

Oreopoulos, J. and C.M. Yip, Combined scanning probe and total internal reflection fluorescence microscopy. Methods, 2008. 46(1): p. 2-10.

See Figure 4.

However, in a very similar instrumental setup, the authors of this paper did not:

Sarkar, A., R.B. Robertson, and J.M. Fernandez, Simultaneous atomic force microscope and fluorescence measurements of protein unfolding using a calibrated evanescent wave. Proceedings of the National Academy of Sciences of the United States of America, 2004. 101(35): p. 12882-12886.

Other papers from the Oheim group that might interest you:

Schapper, F., J.T. Goncalves, and M. Oheim, Fluorescence imaging with two-photon evanescent wave excitation. European Biophysics Journal with Biophysics Letters, 2003. 32(7): p. 635-643.

Oheim, M. and F. Schapper, Non-linear evanescent-field imaging. Journal of Physics D-Applied Physics, 2005. 38(10): p. R185-R197.


Cheers,

John Oreopoulos



On 2016-07-08, at 4:10 PM, Martin Wessendorf wrote:

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> To join, leave or search the confocal microscopy listserv, go to:
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>
> [Posted on behalf of Dr. Alexander Asanov.]
>
> Dear colleagues,
>
> Are you aware of quantitative data for the intensity of stray light in TIRF microscopy? I found only one paper by A. Mattheyses and D. Axelrod, 2006, which quantitatively estimated the intensity of stray light to be ~10–15% relative to the intensity of the evanescent wave at the TIRF interface [Mattheyses, A. L., and Axelrod D. Direct measurement of the evanescent field profile produced by objective-based total internal reflection fluorescence. J Biomed Opt. 2006 Jan-Feb;11(1):014006.]
>
> There are two 2014 papers by Martin Oheim’s group that referenced most of the literature and analyzed sources of stray light, but did not make a quantitative comparison. [Brunstein M, Teremetz M, Hérault K, Tourain C, Oheim M. Eliminating unwanted far-field excitation in objective-type TIRF. Part I. identifying sources of nonevanescent excitation light. Biophys J. 2014 Mar 4;106(5):1020-32.   Brunstein M, Hérault K, Oheim M. Eliminating unwanted far-field excitation in objective-type TIRF. Part II. combined evanescent-wave excitation and supercritical-angle fluorescence detection improves optical sectioning. Biophys J. 2014 Mar 4;106(5):1044-56.]
>
> Does anybody know additional published or unpublished data with Quantitative comparison of the stray light with the evanescent wave?  Thank you.
>
> Best regards,
> Alexander Asanov, Ph.D.
> TIRF Labs
> [hidden email]
>
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